• Title/Summary/Keyword: microbiological medium

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Proteolytic Yeasts Isolated from Mackerel (Scomber japonicus) (고등어에서 분리된 부패성 효모)

  • OH Eun-Gyong;PARK Mi-Yeon;CHANG Dong-Suck
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.4
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    • pp.471-476
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    • 1998
  • Microbiological spoilage of marine fish is complex process occurring by bacteria, yeasts and molds. There have been rare study for saprophytic yeasts although having enormous numbers of bacteriological studies on the spoilage of marine fish. The 14 genera of yeasts isolated from mackerel (Scomber japonicus) with high frequency of occurrence were Candida sp., Rhodotorula sp., Torulopsis sp., Cryptotoccus sp. and Tricosporon sp. Among these ones Candida lipolytica was identified as the strongest proteolytic yeast, then named Candida lipolytica FM5 (C. lipolytica FM5). C. lipolytica FM5 showed optimum growth at $25^{\circ}C$, pH 7.0 and could grow at $5^{\circ}C$ and in medium containing $10\%$ sodium chloride, To evaluate the saprophytic activity of the selected strain, C, lipolytica FM5 and Pseudomonas fluorescens ATCC 17571 which is one of representative spoilage bacteria were individually inoculated into the sterilized fish muscle homogenates, and then pH changes and volatile basic nitrogen (VBN) values were checked during the storage at various temperatures. According to the experimental results, the productions of VBN by C. lipolytica FM5 in the fish muscle homogenates were 50 mg-N/100 g at $5^{\circ}C$, 152 mg-N/100 g at $15^{\circ}C$ and 379 mg-N/100 g at $25^{\circ}C$ for 1 week storage, respectively. Above results were nearly same as in case of Ps. fluorescens ATCC 17571 inoculation. It suggest that sapyophytic yeasts also have important role in spoilage of marine fish.

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The Effect of Moisture Control on Fermentation Characteristics of Barley and Rye Silages (수분 조절이 보리와 호밀 silage의 발효특성에 미치는 영향)

  • Lee, Jong-Chan;Kim, Sam-Churl
    • Journal of agriculture & life science
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    • v.44 no.1
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    • pp.25-32
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    • 2010
  • This study examined the effect of moisture level at ensiling on fermentation characteristics of barley and rye silage. The moisture levels, 60 (low; LM), 70(medium; MM) and 80% (high; HM), were controled by adding water or pre-wilting. Barley silage had higher pH and latate:acetate ratio in LM than the other treatments, but those of rye in MM were higher. The concentrations of lactate, total FA and acetate in HM were higher than the other treatments, but propionate concentration in LM was higher than HM. Total N concentrations of barley and rye were highest in MM and LM, respectively. The $NH_3-N$ concentration and total N:$NH_3-N$ ratio of barley were higher in HM than those in LM and MM. With increasing moisture content, buffering capacity of barley and rye silages increased, whereas decreased by increase of pH. There was a negative correlation between moisture content vs. pH of barley and rye silages. However, moisture content vs. the concentrations of total VFA and $NH_3-N$ and the ratio of total N:$NH_3-N$ had a positive correlation. Tn conclusion, the ideal moisture content of barley and rye for silage was 70-80%, but silage quality could be rapidly decreased by pre-wilting to 60% moisture content.

Isolation and Cultural Characterization of Antibacterial Substance Producing Microbes (항균성 물질 생산 균주의 분리 및 배양학적 특성)

  • Park, Seok-Kyu;Cho, Young-Su;Shon, Mi-Yae;Gal, Sang-Wan;Lee, Sang-Won
    • Food Science and Preservation
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    • v.14 no.2
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    • pp.194-200
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    • 2007
  • In order to enhance the functionality and storage period of traditional fermented foods, the strain CH-14, which To enhance the quality of traditional fermented foods, and to lengthen acceptable storage periods, a bacterial strain, CH-14, showing potent enzyme activities and antibacterial capabilities, was isolated and characterize4 The bacterium wn Gram-positive, catalase-positive, oxidase-negative, formed endospores, expressed flagella, was rod-shaped, and had dimensions of 0.5 0.7m and 3.5 4.2m. The bacterium CH-14 was identified as Bacillus subtilis using Bergey's Manual of Systematic Bacteriology, Bergey's Manual of Determinative Bacteriology, and an API 50 CHL Carbohydrate Test Kit. An optimum growth medium contained 2% (w/v) cellobiose as a carbon source, a mixture of 0.5% (w/v) yeast extract and 0.5% (w/v) peptone as nitrogen sources, and 0.05% (w/v) $MgSO_4{\cdot}7H_2O$. The optimal culture temperature and the optimal initial pH were in the ranges of 30 $45^{\circ}C$ and 4.5 10.0, respectively. Maximum production of the antibacterial substance occurred after 24h of culture. The minimum inhibitory concentrations of the antibacterial substance were 5mg bacterial dry weight/mL against E. coli and P. mirabilis, and 10 mg/mL against S. aureus, S. enteritidis and V. parahaemolyticus.

Removal of mycotoxin ochratoxin A by isolated bacteria (분리세균에 의한 진균독소 ochratoxin A의 제거)

  • Choi, Ho-Yeong;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.55 no.1
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    • pp.33-38
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    • 2019
  • Ochratoxin A (OTA), one of mycotoxins produced mainly by Aspergillus is a common contaminant of stored grains, posing health hazards to human and livestock. The aim of this study is to explore ability of isolated bacteria Bacillus subtilis AF13 and Streptomyces shenzhenensis YR226 to remove OTA. AF13 and YR226 could remove 94.23 and 97.73% of OTA ($100{\mu}g/L$), respectively during 24 h incubation in NB medium. When cultures of two strains were separated into washed cells and cell-free supernatant, the supernatant of both strains removed more than 90% of $100{\mu}g/L$ OTA, and 98.88% of OTA could be also removed by the washed cells of YR226. OTA removal occurred in a few second by the supernatant of both strains, and treatments of autoclaving, proteinase K and chymotrypsin did not affect the OTA removal by the culture supernatants, which indicate that some thermostable and non-proteinaceous substances secreted by these bacteria may be involved in OTA removal in these two bacteria. These results suggest that AF13 and YR226 can be used to remove OTA from OTA-contaminated grains and feeds, and therefore decrease economic damage in agriculture and feed industry.

Inhibition of growth and toxin production of ochratoxigenic Aspergillus spp. by isolated bacteria (분리세균에 의한 ochratoxin 생성 Aspergillus spp.의 생장 및 독소생성 저해)

  • Hwang, Ji-Seon;Choi, Ho-Yeong;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.226-233
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    • 2019
  • Ochratoxin A (OTA) that is one of mycotoxins produced mainly by Aspergillus spp. is a common contaminant of stored grains and poses health hazards to human and livestock. The aim of this study is to explore the ability of isolated bacteria Bacillus subtilis AF13 and Streptomyces shenzhenensis YR226 to inhibit growth and OTA production of 3 ochratoxigenic Aspergillus strains. The antifungal activity against mycelial growth and sporulation of Aspergillus strains was examined by coculture with AF13 and YR226 on potato dextrose agar plate. AF13 and YR226 reduced 77.58 and 78.48% of fungal colony radius, respectively, and both strains inhibited fungal sporulation up to 99% in 10 days of incubation. YR226 also reduced more than 91% of spore germination of 3 fungal strains. When Aspergillus strains were cocultured with AF13 or YR226 in yeast extract sucrose medium, mycelial growth and OTA production decreased in all three fungal strains. In particular, AF13 completely inhibited the mycelial growth of A. alutaceus and inhibited its OTA production by 99%, and YR226 also reduced mycelial growth and toxin production up to 99%, respectively. Antimicrobial substances produced by AF13 and YR226 included siderophore, chitinase, protease, ${\beta}$-1,3-glucanase and biosurfactant. These results suggest that AF13 and YR226 can be used in a biological method to prevent valuable crops against mycotoxigenic fungi, and therefore decrease economic damage in agriculture and feed industry.

Comparative assessment of urban stormwater low impact strategies equipped with pre-treatment zones (침강지 시설이 조성된 LID 시설의 환경적 영향평가)

  • Yano, K.A.V.;Reyes, N.J.D.G.;Jeon, M.S.;Kim, L.H.
    • Journal of Wetlands Research
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    • v.21 no.2
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    • pp.181-190
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    • 2019
  • Recently, Low impact development techniques, a form of nature-based solutions (NBS), were seen cost-efficient alternatives that can be utilized as alternatives for conventional stormwater management practices. This study evaluated the effectiveness of an infiltration trench (IT) and a small constructed wetland (SCW) in treating urban stormwater runoff. Long-term monitoring data were observed to assess the seasonal performance and cite the advantages and disadvantages of utilizing the facilities. Analyses revealed that the IT has reduced performance during the summer season due to higher runoff volumes that exceeded the facility's storage volume capacity and caused the facility to overflow. On the other hand, the pollutant removal efficiency of the SCW was impacted by the winter season as a result of dormant biological activities. Sediment data also indicated that fine and medium sand particles mostly constituted the trapped sediments in the pretreatment and media zones. Sediments in SCW exhibited a lower COD and TN load due to the phytoremediation and microbiological degradation capabilities of the system. This study presented brief comparison LID facilities equipped with pre-treatment zones. The identified factors that can potentially affect the performance of the systems were also beneficial in establishing metrics on the utilization of similar types of nature-based stormwater management practices.

Characteristics of Unrecorded Wild Yeasts Obtained from the Soil of Spices Plant Fields and its Physiological Functionality (향신료 재배 토양으로부터 분리한 국내 미기록 야생효모들의 균학적 특성 및 생리활성)

  • Han, Sang-Min;Kim, Ji-Yoon;Kim, Changmu;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.75-81
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    • 2019
  • The goal of this study was to characterize unrecorded wild yeasts from soils of spices plants fields and further, to elucidate its anti-demential activities and tyrosinase inhibitory activity. Piskurozyma taiwanensis R4-1 (NIBRFGC000502619), Nadsonia starkeyi-henricii R6-2 (NIBRFGC000502618), and Canadida friedrichii M12-6 (NIBRFGC000502615) isolated from soil of garlic field represented newly recorded yeast strains in Korea. Vishniacozyma peneaus I2-9 (NIBRFGC000502617) and Cryptococcus aspenensis I21-1 (NIBRFGC000502616) from soil of ginger field represented also newly recorded yeast strains, and microbiological characteristics of its fifteen yeast strains were investigated. All of these unrecorded yeasts exhibited oval-global shape and have ascospores except Canadida friedrichii M12-6. Piskurozyma taiwanensis R4-1 and Canadida friedrichii M12-6 grew well in vitamin-free medium, and Piskurozyma taiwanensis R4-1 was halotolent growing in 10% NaCl-containing yeast extract peptone dextrose (YPD) broth. After prepared cell-free of the unrecorded wild yeasts, acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities as anti-dementia activity and tyrosinase inhibitory activity as whitening activity were determined. Cell-free extract from Canadida friedrichii M12-6 had the highest tyrosinase inhibitory activity of 14.4%.

Antimicrobial activities of Bacillus subtilis DS660 and Paenibacillus polymyxa DS842 (Bacillus subtilis DS660과 Paenibacillus polymyxa DS842의 항균활성)

  • Lee, Da-Sol;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.354-361
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    • 2018
  • This study was carried out to isolate soil bacteria with antimicrobial activity and evaluate antimicrobial substances produced by isolated bacteria. Among many isolates Bacillus subtilis DS660 and Paenibacillus polymyxa DS842 showed high antimicrobial activities against 6 species of microbial residents on human skin and 3 species of pathogenic bacteria. DS660 and DS842 showed 15.3~26.8 and 11.3~27.5 mm of inhibition zone diameter, respectively on nutrient agar medium against most target bacteria and fungi. DS660 and DS842 produced $57{\pm}8$ and $170{\pm}15{\mu}mol/ml$ of siderophore, respectively as an antimicrobial substance. Analysis of ethyl acetate extract of culture supernatants of DS660 and DS842 suggested production of glycolipid biosurfactant which reduced surface tension of culture supernatant of DS660 and DS842 from 60.0 to 40.3 and 30.3 mN/m, respectively. DS660 and DS842 also showed $169.2{\pm}9.9$ and $357.2{\pm}13.7nmol/min/mg$ protein of ${\beta}-1,3$-glucanase activity, respectively, and hydrolyzed cell wall components of 3 bacterial species. These results suggest that B. subtilis DS660 and P. polymyxa DS842 may be utilized as an environment-friendly biocontrol agent against some skin microbes and pathogenic bacteria.

Substrate chain-length specificities of polyhydroxyalkanoate synthases PhaC1 and PhaC2 from Pseudomonas aeruginosa P-5 (Pseudomonas aeruginosa P-5에 존재하는 polyhydroxyalkanoate synthase PhaC1과 PhaC2의 기질특이성)

  • Woo, Sang Hee;Lee, Sun Hee;Rhee, Young Ha
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.455-462
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    • 2016
  • Pseudomonas aeruginosa P-5 is an unusual organism capable of synthesizing polyhydroxyalkanoates (PHAs) consisting of 3-hydroxyvalerate (3HV) and medium-chain-length (MCL) 3-hydroxyalkanoate (3HA) monomer units when C-odd alkanoic acids are fed as the sole carbon source. Evaluation of the substrate chain-length specificity of two P. aeruginosa P-5 PHA synthases ($PhaC1_{P-5}$ and $PhaC2_{P-5}$) by heterologous expression of $PhaC1_{P-5}$ and $PhaC2_{P-5}$ genes in Pseudomonas putida GPp104 revealed that $PhaC2_{P-5}$ incorporates both 3HV and MCL 3HAs into PHA, whereas $PhaC1_{P-5}$ favors only MCL 3HAs for polymerization. In order to obtain $PhaC2_{P-5}$ mutants with altered substrate specificity, site-specific mutagenesis for $PhaC2_{P-5}$ was conducted. Amino acid substitutions of $PhaC2_{P-5}$ at two positions (Ser326Thr and Gln482Lys) were very effective for synthesizing copolymers with a higher 3HV fraction. When recombinant P. putida GPp104 harboring double mutated $phaC2_{P-5}$ gene ($phaC2_{P-5}QKST$) was grown on nonanoic acid, 2.5-fold increase of copolymer content with 3.8-fold increase of 3HV fraction was observed. The $phaC2_{P-5}QKST$-containing Ralstonia eutropha PHB-4 supplemented with valeric acid also produced copolymers consisting of 3HV and 3-hydroxyheptanoate with a high 3HV fraction. These results suggest that recombinants containing $phaC2_{P-5}QKST$ could be useful for production of new PHA copolymers with improved material properties.

Construction and characterization of heterozygous diploid Escherichia coli (2배체 대장균의 제조와 그 특성)

  • Jung, Hyeim;Lim, Dongbin
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.406-414
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    • 2016
  • Among 6 leu codons, CUG is the most frequently used codon in E. coli. It is recognized by leu-tRNA(CAG) encoded by four genes scattered on two chromosomal loci (leuT and leuPQV ). In the process of constructing a strain with no functional leu-tRNA (CAG) gene on chromosome, we made two mutant strains separately, one on leuPQV locus (${\Delta}leuPQV$), and the other on leuT locus [$leuT^*$(GAG)], where the anticodon of leuT was changed from CAG to GAG, thereby altering its recognition codon from CUG to CUC. We attempted to combine these two mutations by transduction using $leuT^*$(GAG) strain as a donor and ${\Delta}leuPQV$ strain as a recipient. Large and small colonies appeared from this transduction. From PCR and DNA sequencing, large colony was confirmed to be the reciprocal recombinant as expected, but the small colonies contained both mutant $leuT^*$(GAG) and wild type leuT (CAG) genes in the cell. This heterozygous diploid strain did not show any unusual morphology under microscopic observation, but, interestingly, it showed a linear growth curve in rich medium with much slower growth rate than wild type cell. It always formed homogenous small colonies in the selection medium, but, when there was no selection, it readily segregated into $leuT^*$(GAG) and leuT (CAG). From these observations, we suggested that the strain with both $leuT^*$(GAG) and leuT (CAG) genes was not a partial diploid (merodiploid), but a full diploid cell having two different chromosomes. We proposed a model explaining how such a heterozygous diploid cell was formed and how and why its growth showed a linear growth curve.